基于RT-PCR对低温沼气发酵过程中巴氏德醋杆菌的分析  

Analysis of Acetobacter pastcurianum in Low Temperature Biogas Fermentation Process by RT-PCR

作　　者：代金平[1,2] 冯蕾 谢玉清[1,2] 王志方 陈竞[1,2] 王小武 周留艳[1,2] 张慧涛 古丽·艾合买提[1,2] 杨新平 DAI Jinping;FENG Lei;XIE Yuqing;WANG Zhifang;CHEN Jing;WANG Xiawu;ZHOU Liuyan;ZHANG Huitao;GULIahmat;YANG Xinping(Institute of Microbiology,Xinjiang Academy of Agricultural Sciences,Urumqi 830091,China;Xinjiang Key Laboratory of Special Enviromental Microbiology,Urumqi 830091,China;Xinjiang WanKangChengYi fertilizer Co,Ltd,Urumqi 831201,China)

机构地区：[1]新疆农科院微生物应用研究所,新疆乌鲁木齐830091 [2]新疆特殊环境微生物重点实验室,新疆乌鲁木齐830091 [3]新疆万康诚一肥业有限公司,新疆乌鲁木齐831201

出　　处：《中国沼气》2022年第4期31-35,共5页China Biogas

基　　金：自治区科技支疆项目计划项目(2020E0202);新疆维吾尔自治区自然科学基金项目(2021D01B65)。

摘　　要：巴氏德醋杆菌(Acetobacter pastcurianum)在低温沼气发酵过程中能够提高牛羊粪沼气产量,同时也可增加发酵残渣纤维素含量。为探究其在低温沼气发酵过程中的作用,使用荧光定量PCR的方法对不同处理发酵过程中添加的巴氏德醋杆菌(Acetobacter pastcurianum)进行定量分析。试验结果表明,巴氏德醋杆菌(Acetobacter pastcurianum)质粒标准曲线方程为y=-3.024x+30.73,误差为0.0680。处理2、4、7中巴氏德醋杆菌(Acetobacter pastcurianum)基因拷贝数分别为1.83 fg·μL^(-1)、33.77 fg·μL^(-1)和31.93 fg·μL^(-1),巴氏德醋杆菌(Acetobacter pastcurianum)基因拷贝数分别比对照组高出90%、54%和45.6%,且3个处理的产气量也相对较高。建立低温沼气发酵中添加巴氏德醋杆菌的荧光定量检测体系,可以更加快速灵敏地进行添加菌的相关性变化规律分析,为北方地区冬季低温沼气发酵促进菌剂的筛选奠定相关理论基础。In low temperature biogas fermentation process,Acetobacter pastcurianum can effectively increase biogas production of cattle and sheep manure,as well as cellulose content of biogas fermentation residue.In order to explore its role in low-temperature biogas fermentation,fluorescence quantitative PCR was used to quantitatively analyze Acetobacter pasteurianum added in the process of low-temperature fermentation under different treatment conditions in this paper.The results showed that the equation of standard curve is y=-3.024x+30.73 with an error of 0.0680.The concentrations of 16S rDNA of Acetobacter pastcurianum were 41.83 fg·μL^(-1),33.77 fg·μL^(-1) and 31.93 fg·μL^(-1) in the sample 2,4 and 7 respectively.The copy number of Acetobacter pastcurianum gene was 90%,54% and 45.6% higher than that of the control group,respectively,and the gas production of the three treatments was relatively high.Establishing the detection system by real-time fluorescent quantitative PCR can quickly and sensitively quantify Acetobacter pastcurianum more in low-temperature biogas fermentation,meanwhile determine the relevance between the bacteria and gas production.It may lay a theoretical foundation on the screening of microbial inoculums for low temperature biogas fermentation in northern China.

关 键 词：巴氏德醋杆菌 低温 沼气发酵 RT-PCR 绝对定量 

分 类 号：S216.4[农业科学—农业机械化工程] X713[农业科学—农业工程]